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Your Position: Home > Enzymes for In-Depth Antibody Characterization

Enzymes for In-Depth Antibody Characterization

Enzymes for In-Depth Antibody Characterization
Background
Therapeutic antibodies used as biopharmaceuticals are highly complex molecules requiring strict monitoring and control during manufacturing. Due to its nature as a biological drug, drug safety and innate variation in production methods is always a cause for concern. Thus, a series of workflows for comprehensive, in-depth analytical characterization should be applied to analyze and characterize product quality.
With the approval of monoclonal antibody products dramatically increasing each year such as mAbs, Fc-fusions, Fab, antibody-drug conjugates, bispecific mAbs (bsAbs) and bispecific T cell engagers (BiTE), understanding individual post-translational modifications (PTMs) can give a better insight and prediction as to the in vivo efficacy, pharmacokinetics, and mechanism of action of manufactured Abs.
Characterization of Abs is usually performed in several perspectives, starting from top-down analysis (overall structure) to bottom-up analysis (peptide sequencing), and finally, middle-level analysis (Fc, Fv, PTMs).
To enable antibody characterization methods, ACROBiosystems has developed a series of enzymes to assist with the characterization of antibodies and their related PTMs.
Product List
Product DetailsCat. No.Unit Definition
Endo H (500U/μl)ENH-S5116One unit is defined as the amount of enzyme required to remove >95% of the carbohydrate from 10µg of denatured RNase B in 1 hour at 37°C in a total reaction volume of 10µl.
Endo S (200U/μl)ENS-S5143One unit deglycosylates ≥ 95% of 1 μg human IgG, when incubated in 10 mM sodium phosphate, 150 mM NaCl, pH7.4 at 37°C for 30 min.
SpeB (40U/μl)SPB-S5115One unit digests ≥ 95% of 1 µg human IgG1 when incubated in PBS ith 5 mM DTT or TCEP, pH 7.4 at 37°C for 1 hour.
IdeS (20U/μl)IDS-S5143One unit digests ≥ 95% of 1 μg human IgG when incubated in 10 mM sodium phosphate, 137 mM NaCl, 2.7 mM KCl, pH7.4 at 7°C for 30min.
Tools for Glycan Analysis
The structure and type of N-linked glycans can affect the immunogenicity, pharmacokinetics, and pharmacodynamics of therapeutic proteins such as monoclonal antibodies (mAbs) and Fc fusion proteins. Enzymes such as Endo H and Endo S provide type-specific cleavages of N-linked glycans for subsequent analysis.

Endo H (500U/μl) (Cat .No. ENH-S5116)

Application:

Endo H cleaves asparagine-linked (N-linked) hybrid or high mannose oligosaccharides but not complex oligosaccharides.
Cleavage of high mannose or hybrid N-glycans

Endo H (500U/μl) (Cat .No. ENH-S5116)

Product Features:

Selective action: Does not cleave complex glycans
≥95% purity, as determined by SDS-PAGE
Optimal pH:5-6
Reaction time: 1h

Endo S (200U/μl) (Cat .No. ENS-S5143)

Application:

Endo S preferentially cleaves complex oligosaccharides and avoids the cleavage of asparagine-linked (N-linked) hybrid or high mannose oligosaccharides.
Cleaves complex glycans, limited activity on high-mannose and hybrid-type glycans.

Endo S (200U/μl) (Cat .No. ENS-S5143)

Product Features:

Highly efficient IgG-specific endoglycosidase
≥95% purity as determined by SDS-PAGE
Reaction time: 30 min
Antibody Subunit Fragmentation

SpeB (40U/μl) (Cat. No. SPB-S5115)

Application:

SpeB enzyme digests IgG to generate Fab and Fc fragments. Purified and homogenous Fab fragments can be used in many applications, including:
Studying the monovalent binding of Fab fragments to the antigen.
Elimination of Fc-mediated effector functions.
Reduced unspecific binding from Fc interactions.
 

SpeB (40U/μl) (Cat. No. SPB-S5115)

Product Features:

Cleavage site (Human IgG1): KTHT / CPPCPAP (above the hinge)
Reaction time: 60min

IdeS (20U/μl) (Cat. No. IDS-S5143)

Application:

IdeS protease specifically cleaves IgGs below the hinge region into F(ab´)2 and Fc fragments.

IdeS (20U/μl) (Cat. No. IDS-S5143)

Product Features:

Cleavage site (Human IgG1): CPAPELLG / GPSVF (below the hinge)
Reaction time: 30 min
 
ACRO Quality

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